Data from method development and validation studies as predictors of method performance

Problems caused by:

Mobile phase

Column

Sample/standard preparation

Pump

Injector

Detector & data system

Essential tools

Monitoring system performance

Preventative maintenance

Practical exercises:

Revision of topics in the introductory course, plus:

Sample filtration

Priming a dry pump (e.g. after maintenance)

Diagnosing leaks, blockages and pressure fluctuations

HPLC/UHPLC Method Development

A two-day course on method development for liquid chromatography including:

Revision of chromatographic theory

Nature of the analyte(s):

Polarity/solubility

Functionality

Detectability

Stability

Nature of the matrix:

Analyte concentration

Ease of extraction

Chromatographic interference

Method requirements for accuracy, precision and sensitivity

Sample preparation approaches

Developing the separation

Selection of initial conditions

Sample concentration and injection volume

Isocratic and gradient elution

Optimising linearity, sensitivity, selectivity and resolution

Strategies for ionic and non-ionic analytes

Chiral separations

Detector considerations

Biochemical separations

Method validation and transfer

Practical exercises:

Trainees will develop two separations: the first will be for a binary mixture containing a polar molecule and a non-polar species; the second will be for a ternary mixture containing ionic and non-ionic analytes.